A protein's three-dimensional structure is closely related to its biological activity. For protein therapeutics (biologics), their three-dimensional structure, to a certain degree, determine their metabolism (PK/PD) and immunogenicity. These are some of the most important parameters besides their biological activities. Because of the importance of protein 3-D structure, several technologies have been used to analyze the 3-D conformation of biologics during their development. The most common technologies include 1) Molecular Sieve or Gel Filtration; 2) Ultracentrifugation; 3) protein fluorescence; 4) CD spectrum; 5) Non-denature gel electrophoresis. However, there are major limitations from these technologies including: 1) low sensitivity: the result can only provide an overall measurement; it can't distinguish differences in a certain region. 2) The analysis is slow, sometimes more than 24 hours are needed for the analysis. 3) Low throughput: only one or a few samples can be analyzed at a time.
Therefore a more sensitive, accurate and fast method is necessary to provide the 3-dimensional structure or conformation of proteins, especially for the development of biologics.